Genetic diversity and aquaculture conservation for a threatened Neotropical catfish

Due to the ecological importance of Lophiosilurus alexandri, the present work evaluated its genetic representativeness by comparing wild stocks to broodstocks that were kept at three restocking hatcheries along the São Francisco River. A total of 97 samples were genotyped for newly developed microsatellite markers. Low levels of genetic diversity (average alleles number of 4.2 alleles) were detected in all cases, being more severe in captive groups. Significant pairwise FST and DEST values, Structure, and DAPC analyses showed that wild animals were structured in two groups, and a third group was formed by captive animals, evidencing the need to adopt genetic criteria to retain genetic diversity in the hatcheries. For this reason, three full-sib families were constructed to select the best relatedness estimator for L. alexandri and establish a cut-off value aimed to avoid full-sibling matings in the hatcheries. Two estimators, Wang (RW) and Lynch & Li (RLL), were accurate in reflecting the relatedness level for full-sibs in this species. According to them, less than 50% of the potential breeding matings in the three hatcheries are advisable. The innate low diversity of L. alexandri highlights the importance of minimizing inbreeding and retaining genetic diversity towards the species recovery.(AU)

Devido à importância ecológica de Lophiosilurus alexandri, o presente trabalho avaliou sua representatividade genética, comparando estoques selvagens com plantéis de reprodutores de três larviculturas ao longo do Rio São Francisco. Noventa e sete amostras foram genotipadas com marcadores microssatélites recém-desenvolvidos. Baixos níveis de diversidade genética (número médio de alelos de 4,2) foram detectados em todos os casos, sendo mais severo no cativeiro. Os valores de FST e DEST par a par, as análises do Structure e DAPC mostraram a estruturação dos animais selvagens em dois grupos, e um terceiro formado pelas larviculturas, evidenciando a necessidade de adoção de critérios genéticos para retenção da diversidade genética no cativeiro. Por essa razão, três famílias de irmãos completos foram construídas para selecionar o melhor estimador de parentesco para a espécie e estabelecer os valores mínimos de corte para evitar cruzamentos indesejados. Dois estimadores, Wang (RW) e Lynch & Li (RLL), foram eficientes em refletir as relações de parentesco para irmãos completos nessa espécie. Segundo eles, menos de 50% dos potenciais cruzamentos são recomendáveis nas três larviculturas. A baixa diversidade genética inerente ao L. alexandri destaca a importância de minimizar a consanguinidade e evitar perda de diversidade genética, visando a recuperação da espécie.(AU)

Symbiont modulates expression of specific gene categories in Angomonas deanei

Trypanosomatids are parasites that cause disease in humans, animals, and plants. Most are non-pathogenic and some harbor a symbiotic bacterium. Endosymbiosis is part of the evolutionary process of vital cell functions such as respiration and photosynthesis. Angomonas deanei is an example of a symbiont-containing trypanosomatid. In this paper, we sought to investigate how symbionts influence host cells by characterising and comparing the transcriptomes of the symbiont-containing A. deanei (wild type) and the symbiont-free aposymbiotic strains. The comparison revealed that the presence of the symbiont modulates several differentially expressed genes. Empirical analysis of differential gene expression showed that 216 of the 7625 modulated genes were significantly changed. Finally, gene set enrichment analysis revealed that the largest categories of genes that downregulated in the absence of the symbiont were those involved in oxidation-reduction process, ATP hydrolysis coupled proton transport and glycolysis. In contrast, among the upregulated gene categories were those involved in proteolysis, microtubule-based movement, and cellular metabolic process. Our results provide valuable information for dissecting the mechanism of endosymbiosis in A. deanei.

A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni

Schistosomiasis is a major neglected tropical disease caused by trematodes from the genus Schistosoma. Because schistosomes exhibit a complex life cycle and numerous mechanisms for regulating gene expression, it is believed that spliced leader (SL) trans-splicing could play an important role in the biology of these parasites. The purpose of this study was to investigate the function of trans-splicing in Schistosoma mansoni through analysis of genes that may be regulated by this mechanism and via silencing SL-containing transcripts through RNA interference. Here, we report our analysis of SL transcript-enriched cDNA libraries from different S. mansoni life stages. Our results show that the trans-splicing mechanism is apparently not associated with specific genes, subcellular localisations or life stages. In cross-species comparisons, even though the sets of genes that are subject to SL trans-splicing regulation appear to differ between organisms, several commonly shared orthologues were observed. Knockdown of trans-spliced transcripts in sporocysts resulted in a systemic reduction of the expression levels of all tested trans-spliced transcripts; however, the only phenotypic effect observed was diminished larval size. Further studies involving the findings from this work will provide new insights into the role of trans-splicing in the biology of S. mansoni and other organisms. All Expressed Sequence Tags generated in this study were submitted to dbEST as five different libraries. The accessions for each library and for the individual sequences are as follows: (i) adult worms of mixed sexes (LIBEST_027999: JZ139310 - JZ139779), (ii) female adult worms (LIBEST_028000: JZ139780 - JZ140379), (iii) male adult worms (LIBEST_028001: JZ140380 - JZ141002), (iv) eggs (LIBEST_028002: JZ141003 - JZ141497) and (v) schistosomula (LIBEST_028003: JZ141498 - JZ141974).

Evaluation of window cohabitation of DNA sequencing errors and lowest PHRED quality values

When analyzing sequencing reads, it is important to distinguish between putative correct and wrong bases. An open question is how a PHRED quality value is capable of identifying the miscalled bases and if there is a quality cutoff that allows mapping of most errors. Considering the fact that a low quality value does not necessarily indicate a miscalled position, we decided to investigate if window-based analyses of quality values might better predict errors. There are many reasons to look for a perfect window in DNA sequences, such as when using SAGE technique, looking for BLAST seeding and clustering sequences. Thus, we set out to find a quality cutoff value that would distinguish non-perfect windows from perfect ones. We produced and compared 846 reads of pUC18 with the published pUC consensus, by local alignment. We then generated a database containing all mismatches, insertions and gaps in order to map real perfect windows. An investigation was made to find the potential to predict perfect windows when all bases in the window show quality values over a given cutoff. We conclude that, in window-based applications, a PHRED quality value cutoff of 7 masks most of the errors without masking real correct windows. We suggest that the putative wrong bases be indicated in lower case, increasing the information on the sequence databases without increasing the size the files.

Clustering of Schistosoma mansoni mRNA sequences and analysis of the most transcribed genes: implications in metabolism and biology of different developmental stages

The study of the Schistosoma mansoni genome, one of the etiologic agents of human schistosomiasis, is essential for a better understanding of the biology and development of this parasite. In order to get an overview of all S. mansoni catalogued gene sequences, we performed a clustering analysis of the parasite mRNA sequences available in public databases. This was made using softwares PHRAP and CAP3. The consensus sequences, generated after the alignment of cluster constituent sequences, allowed the identification by database homology searches of the most expressed genes in the worm. We analyzed these genes and looked for a correlation between their high expression and parasite metabolism and biology. We observed that the majority of these genes is related to the maintenance of basic cell functions, encoding genes whose products are related to the cytoskeleton, intracellular transport and energy metabolism. Evidences are presented here that genes for aerobic energy metabolism are expressed in all the developmental stages analyzed. Some of the most expressed genes could not be identified by homology searches and may have some specific functions in the parasite